Increasing the effective blood level of isoniazide



United States Patent Uni-ice 3,038,837 Patented June 12, 1962 The present invention relates to a method of increasing the tuberculostatically effective blood level of isonicotinic acid hydrazide and to compositions for achieving said effect.

Isonicotinic acid hydrazide is, at present, one of the most effective tuberculostatic agents being used in the therapy of human tuberculosis. It is usually administered to patients in doses of to mg. per kg. of body Weight in 3 subdivided doses per day. The blood and tissue levels of tuberculostatically effective isonicotinic hydrazide thus obtained, however, vary considerably with different patients. This is apparently due to the presence of various types of inactivators (see Roger S. Mitchell and J. Caroll Bell, Modern Chemotherapy of Tuberculsosis, published by Medical Encyclopedia, Inc., New York, 1958). The presence of fast inactivators might result in a very rapid change of the administered isonicotinic acid hydrazide into tuberculostatically ineffective metabolites so that the therapeutical effect of the isonicotinic acid hydrazide treatment become uncertain.

It is one object of the present invention to provide a simple and advantageous method of increasing the tuberculostatically effective blood level of isonicotinic hydrazide.

Another object of the present invention is to provide a composition containing isonicotinic acid hydrazide which composition is capable of increasing its tuberculostatically effective blood level.

These and other objects of the present invention and advantageous features thereof will become apparent as the description proceeds.

According to the present invention it has been found that the tuberculostatically effective isonicotinic acid hydrazide levels can be increased very considerably, especially in humans with inactivators of the fast-type," by administering a mixture which contains N-acetyl-D-glucosaminyl-N-isonicotinic acid hydrazide in addition to isonicotinic acid hydrazide.

Said N-acetyl-D-glucosaminyl isonicotinic acid hydrazide corresponds to the formula In the following specification the isonicotinic acid hydrazide will be indicated, for brevitys sake, as INH and the N-acetyl-D-glucosarninyl-N-isonicotinic acid hydrazide as NAGINH.

On administering a single dose of 4 rug/kg. of INH to patients having an inactivator of the type of fast INH inactivators and determining the INH blood level by using conventional chemical and biological test methods, the following values (Table l) were found:

TABLE 1 Determination of the tuberculostatically effective plasma level of INH and of the sum of INH and its metabolites according to the methods of Nielsch Chemikerzeitung," vol. 82, page 329 (1958), and of Nielsch and Giefer Arzneimittelforschung, vol. 8, in print (1959), after administration of a single dose of 4 mg./l g. of INH as a function of time. Total dose administered: 300 mg.

Free Hours after Sum total tuberculo- Time of day adminiscalculated statically nation as INH in effective gJcc. INH in ug/cc.

The fact that the plasma blood level of free, tuberculostatically effective INH is lower than 0.2 ,ug/cc. after 4 hours proves that the test person contains a fast INH inactivator.

The same test person was given 300 mg. of INH and 750 mg. of NAGINH at the same time. The results are illustrated in Table 2.

TABLE 2 Determination of the tuberculostatically effective plasma level of INH and of the sum total of INH and NAGINH, and their metabolites after a single administration of 4 mg./kg. of INH and 10 rug/kg. of NAGINH.

Total dose administered: 300 mg. of'INH and 750 mg. of NAGINH.

Free Hours after Sum total tuberculo- Time of day adminiscalculated statically nation as INH in effective g./cc. INH in gJce.

The results of these tests show clearly that the blood plasma levels of free tuberculostatically effective INH are substantially higher at all times and that even after 6 hours said level is significantly higher than that achieved by administration of the same amount of INH alone. 'It is evident that the effectiveness of INH in combination with NAGINH lasts about 50% longer than that of INH alone.

To prove that NAGINH alone does not produce a substantial tuberculostatic plasma level in the same test person, and that the results of Table 2 are based on the unexpected effect of the combination of INH and NAGINH, 750 mg. of NAGINH alone were administered to the same test person. The blood plasma levels are given in Table 3.

TABLE 3 Determination of the tuberculostatically effective plasma level of INH according to biological methods, and of the sum total of NAGINH and its metabolites accordsame test person.

J ing to chemical methods after administration of a single dose of 10 mg./kg. of NAGINH.

Total dose: 750 mg. of NAGINH.

Free tuber- Hours Sum total calcuculostatically Time of day after lated as IN H in effective IN H adminisgJcc. (chemiin ,flgJCG. (biotration cally determined) logically determined) 9. 999999? ooooo i-u-nooouooooooalowm TABLE 4 Determination of the tuberculostatically effective plasma level of INH and of the sum total of INH and its metabolites after administering three times 200 mg. each of INH to humans.

Table 4 clearly proves that on conventionally administering INH in three subdivided doses, the tuberculostati cally effective level of INH drops so far overnight that, the next morning, no more tuberculostatic effect is determinable in the plasma and the tissues.

100 mg. of INH were administered in combination with 250 mg. of NAGINH three times daily. The results are given in Table 5.

TABLE 5 Determination of the tuberculostatically effective plasma level of INH and of the sum total of INH, NAGINH and their metabolites after administering three times 100 mg. of INH in combination with 250 mg. of NAGINH daily to humans.

Free tuberculostatically efiective INH in gJcc.

Sum total calculated as IN H in Time of day ngJCG.

Table 5 proves that, when administering three times daily each time 100 mg. of INH only in combination with 250 mg. of NAGINH the blood level of tuberculostatically effective INH is several times higher than when administering twice the amount of INH in three subdivided doses and that there is still exerted a substantial tuberculostatic efiect on the next morning, i.e., the combination produces the required blood levels for a considerably more prolonged period of time than INH.

The above given results were confirmed by extensive pharmacological tests and showed that both biological and chemical methods of determining the INH blood level provided comparable results also with other test persons. Clinical tests also proved the superior therapeutic effect of the new combination.

The above described tests were carried out with mixtures containing isonicotinic acid hydrazide and N-acetyl- D-glucosaminyl-N-isonicotinic acid hydrazide in the proportion of 0.4: 1.0. Although this proportion has proved to be especially effective, the amount of N-acetyl-D-glucosaminyl-N-isonicotinic acid hydrazide may also be higher or even lower. However, a proportion of l.0:l.0 of isonicotinic acid hydrazide to N-acetyl-D-glucosaminyl- N-isonicotinic acid hydrazide should not be exceeded. N-acetyl-D-glucosaminyl-N-isonicotinic acid hydrazide is prepared by heating on the water bath 28 g. of isonicotinic acid hydrazide with 44 g. of N-acetyl-D-glucosamine in a mixture of 200 cc. of ethanol, cc. of water, and 20 cc. of glacial acetic acid for a short period of time, until a clear solution has formed. About 210 cc. of solvent are then distilled off in a vacuum. On standing overnight, crystals separate from the oily residue...

On triturating, the reaction product solidifies to a paste which, on triturating with ethanol, yields an almost pure compound. The reaction product is obtained in the form of its dihydrate. It can be purified by dissolving it in a small amount of cold water and gradually adding acetone to the solution whereby it is precipitated in a substantially pure form. Melting point: C. with decomposition.

Analysis.-C H O -N.2H O; Molecular weight 376.4. Calculated: 44.67% C; 6.43% H; 14.89% N. Found: 44.65% C; 6.42% H; 14.91% N.

The dihydrate is soluble in hot ethanol. On cooling the solution and scratching with a glass rod, the Water-free N-acetyl-D-glucosaminyl-N-isonicotinic acid hydrazide of the melting point 208 C. with decomposition precipitates. Yield: 60 g. corresponding to 80% of the theoretical yield.

Analysis.C H O N molecular weight: 340.3. Calculated: 16.46% N. Found: 16.36% N.

The compositions according to the present invention may be of any desired form, such as in the form of tablets, dr-agees, pills, powders, and the like solid preparations or in the form of solutions, emulsions, suspensions, and the like liquid preparations whereby various diluents may be employed.

In the case of powders, fine uniform dispersion of the two compounds together with a conventional carrier throughout said powder is of importance. Such a fine dispersion can be achieved by intimately mixing and milling the compounds, for instance, in a ball mill with a solid pulverulent extending agent to the desired degree of fineness or by impregnating the already milled, finely powdered, solid carrier with a mixture of the two compounds in water or with a solution thereof in a suitable organic solvent and then removing water or solvent.

A fine dispersion of the two compounds in water, sirup and the like can also be brought about by emulsifying said compounds with the aid of a dispersing or emulsifying agent.

When preparing tablets, pills, powders, and the like solid preparations, the commonly used carriers and diluting agents, binders, and the like tableting adjuvants are employed, such as sugar, lactose, starch, pectin, bolus alba, stearic acid, magnesium stearate, and as binders, gelatin, gum arabic, methyl cellulose, sodium carboxy methyl cellulose yeast extract, agar, tragacanth, and others. It is, of course, understood that any of the tableting materials conventionally used in pharmaceutical practice can be em ployed provided there is no incompatibility with the two compounds.

The content of the two compounds in compositions according to the present invention may vary. Ordinarily, the compositions according to the invention should not contain less than about 25% of the two compounds. The preferred amounts to be employed are between 75% and about 95%. To use greater amounts is also possible although administration of suitable doses becomes more difiicult. Tablets, each containing, for instance, between about 10 mg. and about 100 mg. of isonicotinic acid hydrazide and between about 25 mg. and about 250 mg. of N- acetyl--D-glucosaminyl-N-isonicotinie acid hydrazide have proved to be especially suitable.

The following examples serve to illustrate compositions according to the present invention without, however, being limited thereto.

Example 1 20 g. of isonicotinic acid hydrazide and 50 g. of N- acetyl-D-glusosaminyl-N-isoniootinic acid hydrazide are intimately mixed with 60 g. of wheat starch and 60 g. of lactose and the mixture is granulated. The granulated mixture is mixed with 5 g. of talc powder and 5 g. of stearic acid. The resulting granules are tableted yielding about 1000 tablets of about 200 mg; each tablet containing 20 mg. of isonicotinic acid hydrazide and 50 mg. of N-acetyl-D-glucosaminyl-N-isonicotinic acid hydrazide.

Example 2 A mixture of 50 g. of isonicotinic acid hydrazide and 125 g. of N-acetylaD-glucosaminyl-N-isonicotinic acid hydrazide is mixed with 40 g. of starch and 25 g. of lactose and is granulated by means of a starch paste. 10 g. of talc are added thereto and 1000 pills are pressed from said mixture. The pills are then converted into sugarcoated dragees by means of 50 g. of sugar coating. Each dragee weighs about 300 mg. and contains 50 mg. of isonicotinic acid hydrazide and 125 mg. of N-acetyl-D- glucosaminyl-N-isonicotinic acid hydnazide. Of course, many changes and variations in the composition of the pharmaceutical preparations according to the present invention, in the proportion of isonicotinic acid hydrazide and N-acetyl-D-glucosaminylaN-isonicotinic acid hydr-azide, in the amounts of said compounds present in the preparation, in the manner of preparing the compositions, in the method of using the compositions in therapy and the dosage employed, and the like may be made by those skilled in the art in accordance with the principles set forth herein and in the claims annexed hereto.

It is, of course, understood that the racemic N-acetyl glucosaminyl isonicotinic acid hydrazide may be used as blood level increasing agent although at least twice the amount of the D-isomer is required to produce the desired effect. The L-isomer may also be employed but it requires much larger amounts than are necessary when using the D- isomer.

Although the present invention has proved of great value in human therapy, it may, of course, also be used in veterinary medicine, for instance, in the treatment of tubercular co-ws whereby similar advantageous results are achieved.

I claim:

1. The method of increasing the tuberculostatically efiective blood level of isonicotinic acid hydrazide in humans and animals, said method comprising orally administering a mixture of 10 parts to 100 parts by Weight of isonicotinic acid hydrazide with 25 parts to 250 parts by weight of N-acetyl-D-glucosaminyl-N-isonicotinic acid hydrazide.

2. A pharmaceutical composition for increasing the tuberculostatically effective blood level of isonicotinic acid hydrazide in humans and animals, said composition containing a mixture of 10 parts to 100 parts by weight of isonicotinic acid hydrazide with 25 parts to 250 parts by weight of N-acetyl-D-glucosaminyl-N-isonicotinic acid hydrazide.

References Cited in the file of this patent UNITED STATES PATENTS 2,937,116 Johnson May 17, 1960 2,971,887 Johnson Feb. 14, 1961 OTHER REFERENCES Chemical Abstracts, vol. 48, p. 10230 1954 (abstract of Valdecasas et al., Arch. Inst. Farmacol, exptl. (Madrid) 5, 47-50,1953).

Bernstein et al.; Am. Rev. of Tuberculosis, 67:3, pp. 354 and 365.

Johnson: Nature, 17414433, pp. 744-745, Oct. 16, 1954.

Kass et 31.: Ann. Int. Med., 47:4, pp. 744-745, 758-761, October 1957.

Prescott et al.: P.S.E.B.M., :4, pp. 687-690, Aug.- Sept. 1957.

Prescott et a1.: Antibiotics and Chemotherapy, VIII: 7, pp. 349-353, July 1958. 

2. A PHARMACEUTICAL COMPOSITION FOR INCREASING THE TUBERCULOSTATICALLY EFFECTIVE BLOOD LEVEL OF ISONICOTINIC ACID HYDRAZIDE IN HUMAN AND ANIMALS, SAID COMPOSITION CONTAINING A MIXTURE OF 10 PARTS TO 100 PARTS BY WEIGHT OF ISONICOTINIC ACID HYDRAZIDE WITH 25 PARTS TO 250 PARTS BY WEIGHT OF N-ACETYL-D-GLUCOSAMINYL-N-ISONICOTINIC ACID HYDRAZIDE. 